CN102177959B - Seaweed protein bacteriostatic agent and application of the seaweed protein bacteriostatic agent - Google Patents
Seaweed protein bacteriostatic agent and application of the seaweed protein bacteriostatic agent Download PDFInfo
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- CN102177959B CN102177959B CN2011100485742A CN201110048574A CN102177959B CN 102177959 B CN102177959 B CN 102177959B CN 2011100485742 A CN2011100485742 A CN 2011100485742A CN 201110048574 A CN201110048574 A CN 201110048574A CN 102177959 B CN102177959 B CN 102177959B
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- sargassum protein
- phosphate buffer
- sargassum
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Abstract
The invention relates to the technical field of food safety treatment and particularly provides a seaweed protein bacteriostatic agent. The seaweed protein bacteriostatic agent is freeze-dried powder which is obtained by mixing, freezing and drying the following materials in percent by weight: 1-50% of seaweed protein concentrated solution, 1-5% of sucrose and trehalose, and the balance of phosphate buffer. The invention also provides an application of the seaweed protein bacteriostatic agent in the preservation of red bayberry. The seaweed protein bacteriostatic agent has the characteristics of being convenient and fast to store and transport, convenient to use and environment-friendly; and an experiment shows that the adoption of the seaweed protein bacteriostatic agent prolongs the low-temperature storage period of the red bayberry from original 5-7 days to 15-20 days.
Description
Technical field
The present invention relates to the food security processing technology field, relate in particular to a kind of red bayberry preservation and freshness Sargassum protein inhibiting-bacteria preparation and application thereof of natural and safe.
Background technology
Red bayberry is the special product fruit of China, and fresh fruit is bright in colour, and sour and sweet palatability is with rich flavor, is rich in nutritional labelings such as multi mineral prime element, vitamin, amino acid.Yet Waxberry fruit does not have exocarp and encapsulates, and cedductor is tender, and the Yangtze River Delta Area maturation is high temperature and rainy season, normal temperature shelf life only 2 ~ 3 days, and it is serious to adopt the back loss, sells for the strange land of red bayberry fresh fruit to export with fresh fruit to have brought great puzzlement.In recent years; Application along with foam preservative case; Red bayberry has been realized the medium and long distance sale; Long mycocriny is mashed but the temperature fluctuation in the transporting procedures very easily causes the red bayberry surface, and the chemical bacteriostatic agent that tradition is used is easy to generate potential safety hazard, and uses chemical bacteriostatic agent can bring such as stubborn problems such as residues of pesticides toxicity, increase environmental pollution and phytopathogen develop immunity to drugs for a long time.
The initiative of Environmental compatibility bacteriostatic agent is the inexorable trend of bacteriostatic agent development.Wherein, Obtain from the nature biotechnology clock the inhibited natural products of causal organism directly is used for controlling plant diseases; Or carry out bionical syntheticly according to the chemical constitution of natural products, with controlling plant diseases, be an effective way of Environmental compatibility bacteriostatic agent development.Marine alga is one type of important biology in the ocean, and the marine alga active material is one of focus of marine biology research, and research shows can obtain multiple material with antitumor, anti-oxidant, antibacterium, antimycotic, antiviral activity from marine alga.Therefore, from marine alga, obtain natural products, directly be used for the antibacterial mildew-resistant of agricultural products storage and preservation, more and more receive people's attention with the various pathogens of inhibition or desinsection.
Preserving fruit and vegetable utilizing mainly is master's tradition storage pattern with common low temperature and chemical preservation at present.Such as, publication number is realized the fresh-keeping of Waxberry fruit for the CN101003767A Chinese invention patent provides a kind of liquid for washing fruit of waxberry; Publication number is processed the slowly-releasing parcel and is used for the preservation and freshness red bayberry for the CN1513334A Chinese invention patent provides a kind of 10～30%ClO2 powder that in starch, mixes, and these methods are mainly chemical means, all have the possibility of secondary pollution.Therefore, press for cheap, the efficient and safe more fruits and vegetable stock and preserving freshness means of development, with the ripening and senescence of controlling fruits and vegetables and the edible safety of guaranteeing fruits and vegetables.
Summary of the invention
The object of the invention is; Mashed to the long mycocriny in surface in the red bayberry transporting procedures; The traditional chemical bacteriostatic agent is easy to generate potential safety hazard; And use chemical bacteriostatic agent can bring such as residues of pesticides toxicity, increase stubborn problems such as environmental pollution and phytopathogen develop immunity to drugs etc. for a long time, and provide a kind of technology simple and easy to do Sargassum protein inhibiting-bacteria preparation and application, make the Sargassum protein inhibiting-bacteria preparation that provides can effectively suppress the long bacterium in surface in the red bayberry transporting procedures; Make its low temperature storing and transporting phase extend to 15 ~ 20 days, be fit to vast red bayberry large plantation family and trafficking family and apply by original 5 ~ 7 days.
For realizing goal of the invention of the present invention, the inventor provides following technical scheme:
A kind of Sargassum protein inhibiting-bacteria preparation, it is for being mixed by following weight percentages after the freeze dried powder that obtains is handled in freeze drying:
Sargassum protein concentrate 1～50%, sucrose and trehalose 1～5%, surplus is a phosphate buffer.
The inventor discovers, according to the freeze dried powder that above-mentioned prescription makes, is principal component with the Sargassum protein, is aided with sucrose and trehalose as protein stabiliser, has obtained a kind of natural bacteriostatic preparation that is applicable to Waxberry preservation.Sargassum protein inhibiting-bacteria preparation of the present invention need place-10 to-25 ℃ of low-temperature preservations, to guarantee the activity of Sargassum protein.
As preferably, according to Sargassum protein inhibiting-bacteria preparation of the present invention, wherein, described Sargassum protein concentrate be fresh sheep dwell dish through the phosphate buffer lixiviate, filter, add solid (NH
4)
2SO
4Deposition, dialysis and PEG2000 concentrate and make.The technological means of using in the purge process of Sargassum protein of the present invention gets final product by this area universal method.
As preferably, according to Sargassum protein inhibiting-bacteria preparation of the present invention, wherein, described Sargassum protein concentrate obtains as follows:
(1) break process: get the fresh sheep dish of dwelling, add ice cube and phosphate buffer and carry out fragmentation, dwell dish and phosphate buffer of sheep is the mass ratio of 1:5～20,
(2) lixiviate, filtration, deposition, dialysis and concentration: the product after the break process is in 4 ℃ of following lixiviate 2h; With the coarse filtration of Sargassum protein leaching liquor, remove slag and get filtrating then, the protein extract after the coarse filtration is centrifugal in refrigerated centrifuge; Get supernatant, add solid (NH
4)
2SO
4Get 50% (NH
4)
2SO
4Deposition; Get the phosphate buffer dissolution precipitation and pack in the bag filter, dialyse in the phosphate buffer, each dialysis time>=6h changes liquid number of times>=4 time; After dialysis is accomplished, concentrate with PEG20000 again, obtain the Sargassum protein concentrate.
50% (NH among the present invention
4)
2SO
4Deposition is meant in the supernatant that obtains to add solid (NH
4)
2SO
4Reach 50% saturation degree, leave standstill 6～12h, again at 4 ℃ of albumen precipitations that obtain with the centrifugal 25min of 11000rpm 50% (NH
4)
2SO
4Deposition.
As preferably, according to Sargassum protein inhibiting-bacteria preparation of the present invention, wherein, described sucrose and trehalose are by weight being the 1:1 proportioning.
As preferably, according to Sargassum protein inhibiting-bacteria preparation of the present invention, wherein, described phosphate buffer is the phosphate buffer of the 0.06mol/L of pH 7.5.Experiment shows that Sargassum protein stability in this buffer solution is better.
As preferably, according to Sargassum protein inhibiting-bacteria preparation of the present invention, wherein, the addition of described ice cube is as the criterion between 2～6 ℃ with the temperature in the control shattering process.In shattering process, control temperature, to prevent the Sargassum protein inactivation, experiment shows that 2～6 ℃ of following extractions can not destroy the activity of Sargassum protein.
As preferably, according to Sargassum protein inhibiting-bacteria preparation of the present invention, wherein, described Sargassum protein concentrate is preserved subsequent use down in-25 ℃～-10 ℃.Can realize the low-temperature protection of Sargassum protein.
As preferably, according to Sargassum protein inhibiting-bacteria preparation of the present invention, wherein, during described freeze drying was handled: the cold-trap precooling was to-35 ℃, and vacuum is 30～60Pa, and the time that freeze drying is handled is 8～9h.Can better preserve Sargassum protein.
The present invention also provides the above-mentioned application of Sargassum protein inhibiting-bacteria preparation in Waxberry preservation.
As preferably, according to the application of Sargassum protein inhibiting-bacteria preparation of the present invention in Waxberry preservation, wherein, described application is operated as follows:
Sargassum protein inhibiting-bacteria preparation 1g is dissolved in the 500-2500ml pure water, is sprayed on then on the new arbutus and gets final product.Rationally control Sargassum protein concentration is about 0.02mg/ml, can realize the sterilization purpose.
Compared with prior art, the present invention has following advantage:
1, the invention provides a kind of Sargassum protein inhibiting-bacteria preparation freeze-dried powder preparation, greatly facilitate storage and transportation, and made things convenient for the red bayberry large plantation family and through the use of cancellation.
2, the present invention uses natural seaweed albumen inhibiting-bacteria preparation; The fruit that does not have skin for the no shell of red bayberry and so on; Not only can its storage fresh-keeping phase of effective antibacterial prolongation, and can reduce the pesticide residual contamination of traditional chemical medicament, make the edible red bayberry fresh fruit that more consumers can be more relieved.
3, the present invention has improved the method that use is soaked in traditional bacteriostasis, preservation agent; Not only reduced the use amount of bacteriostatic agent; Reduced the dry link of soaking the fresh-keeping red bayberry in back simultaneously; So not only reduced the red bayberry high temperature exposure time, improved the red bayberry preservation and freshness time accordingly, also reduced of the loss of links such as immersion simultaneously raw material.
Description of drawings
Fig. 1 is Sargassum protein inhibiting-bacteria preparation of the present invention fungistatic effect experimental curve diagram in the red bayberry preservation and freshness.
The specific embodiment
Below in conjunction with embodiment, content of the present invention is described more specifically.Should be appreciated that enforcement of the present invention is not limited to following embodiment, all will fall into protection domain of the present invention any pro forma accommodation and/or the change that the present invention made.
In the present invention, if not refer in particular to, all part, percentages are unit of weight, and all equipment and raw material etc. all can be buied from market or the industry is commonly used.Do not specialize if having, the method that embodiment adopts is this area current techique.
The preparation method of Sargassum protein inhibiting-bacteria preparation, carry out as follows:
(1) preparation of Sargassum protein concentrate:
Get fresh sheep dwell dish put into juice extractor and add an amount of ice cube and the 0.06mol/L phosphate buffer of pH 7.5 carry out fragmentation (shattering process note control temperature at 2 ~ 6 ℃; Prevent the Sargassum protein inactivation), dwell dish raw material and phosphate buffer of sheep is the part by weight of 1:5.After the break process, put into 4 ℃ of refrigerator lixiviate 2h.Use clean one deck gauze with the coarse filtration of Sargassum protein leaching liquor then, remove slag and get filtrating.With the centrifugal 6min in 4 ℃, the refrigerated centrifuge of 6000r/min of the protein extract after the coarse filtration, get supernatant and add solid (NH
4)
2SO
4Reach 50% saturation degree, leave standstill 6h, obtain 50% (NH at 4 ℃ with the centrifugal 25min of 11000rpm again
4)
2SO
4Albumen precipitation.The take a morsel 0.06mol/L phosphate buffer dissolution precipitation of pH 7.5 in the bag filter of packing into, is dialysed in the 0.06mol/L phosphate buffer of pH 7.5, and the 6h that at every turn dialyses changes liquid 5 times.After dialysis is accomplished, concentrate with polyethylene glycol PEG20000, the Sargassum protein concentrate that obtains is preserved subsequent use down in-10 ℃ again.
(2) inhibiting-bacteria preparation preparation and storage:
0.06mol/L phosphate buffer=1%:1%:98% of Sargassum protein concentrate: sucrose and trehalose: pH 7.5 by mass percentage, the weight ratio=1:1 of sucrose and trehalose gets the raw materials ready.
A. above-mentioned raw materials is mixed,
B. can: with the protein solution sterile filling for preparing in cillin bottle,
C. freeze-drying: carry out freeze drying with cryogenic vacuum equipment, vacuumize, jump a queue, capping gets Sargassum protein inhibiting-bacteria preparation finished product, and wherein: the cold-trap precooling is to-35 ℃, and vacuum is 30Pa, and the time that freeze drying is handled is 8h.
D. store: finished product places-10 ℃ of low-temperature preservations.
The preparation method of Sargassum protein inhibiting-bacteria preparation, carry out as follows:
(1) preparation of Sargassum protein concentrate:
Get fresh sheep dwell dish put into juice extractor and add an amount of ice cube and the 0.06mol/L phosphate buffer of pH 7.5 carry out fragmentation (shattering process note control temperature at 2 ~ 6 ℃; Prevent the Sargassum protein inactivation), the 0.06mol/L phosphate buffer of raw material and pH 7.5 is the ratio of 1:10.After the fragmentation, put into 4 ℃ of refrigerator lixiviate 2h., remove slag and get filtrating the coarse filtration of Sargassum protein leaching liquor with clean one deck gauze.At 4 ℃, centrifugal 6min in the refrigerated centrifuge of 6000r/min gets supernatant and adds solid (NH with the protein extract after the coarse filtration
4)
2SO
4Reach 50% saturation degree, leave standstill 12h, obtain 50% (NH at 4 ℃ with the centrifugal 25min of 11000rpm again
4)
2SO
4Albumen precipitation.The take a morsel 0.06mol/L phosphate buffer dissolution precipitation of pH 7.5 in the bag filter of packing into, is dialysed in the 0.06mol/L phosphate buffer of pH 7.5, and the 12h that at every turn dialyses changes liquid 4 times.After dialysis is accomplished, concentrate with polyethylene glycol PEG20000, the Sargassum protein concentrate that obtains is preserved subsequent use down in-20 ℃ again.
(2) inhibiting-bacteria preparation preparation and storage:
0.06mol/L phosphate buffer=50%:5%:45% of Sargassum protein concentrate: sucrose and trehalose: pH 7.5 by mass percentage, the weight ratio=1:1 of sucrose and trehalose gets the raw materials ready.
A. above-mentioned raw materials is mixed,
B. can: with the protein solution sterile filling for preparing in cillin bottle,
C. freeze-drying: carry out freeze drying with cryogenic vacuum equipment, vacuumize, jump a queue, capping gets Sargassum protein inhibiting-bacteria preparation finished product, and wherein: the cold-trap precooling is to-35 ℃, and vacuum is 60Pa, and the time that freeze drying is handled is 9h.
D. store: finished product places-19 ℃ of low-temperature preservations.
Embodiment 3
The preparation method of Sargassum protein inhibiting-bacteria preparation, carry out as follows:
(1) preparation of Sargassum protein concentrate:
Get fresh sheep dwell dish put into juice extractor and add an amount of ice cube and the 0.06mol/L phosphate buffer of pH 7.5 carry out fragmentation (shattering process note control temperature at 2 ~ 6 ℃; Prevent the Sargassum protein inactivation), the 0.06mol/L phosphate buffer of raw material and pH 7.5 is the ratio of 1:20.After the fragmentation, put into 4 ℃ of refrigerator lixiviate 2h., remove slag and get filtrating the coarse filtration of Sargassum protein leaching liquor with clean one deck gauze.At 4 ℃, centrifugal 6min in the refrigerated centrifuge of 6000r/min gets supernatant and adds solid (NH with the protein extract after the coarse filtration
4)
2SO
4Reach 50% saturation degree, leave standstill 10h, obtain 50% (NH at 4 ℃ with the centrifugal 25min of 11000rpm again
4)
2SO
4Albumen precipitation.The take a morsel 0.06mol/L phosphate buffer dissolution precipitation of pH 7.5 in the bag filter of packing into, is dialysed in the 0.06mol/L phosphate buffer of pH 7.5, and the 8h that at every turn dialyses changes liquid 6 times, to remove (NH as much as possible
4)
2SO
4After dialysis is accomplished, concentrate with polyethylene glycol PEG20000, the Sargassum protein concentrate that obtains is preserved subsequent use down in-20 ℃ again.
(2) inhibiting-bacteria preparation preparation and storage:
0.06mol/L phosphate buffer=10%:2%:88% of Sargassum protein concentrate: sucrose and trehalose: pH 7.5 by mass percentage, the weight ratio=1:1 of sucrose and trehalose gets the raw materials ready.
A. above-mentioned raw materials is mixed,
B. can: with the protein solution sterile filling for preparing in cillin bottle,
C. freeze-drying: carry out freeze drying with cryogenic vacuum equipment, vacuumize, jump a queue, capping gets Sargassum protein inhibiting-bacteria preparation finished product, and wherein: the cold-trap precooling is to-35 ℃, and vacuum is 50Pa, and the time that freeze drying is handled is 8h.
D. store: finished product places-25 ℃ of low-temperature preservations.
Experimental example 1The stability experiment of Sargassum protein inhibiting-bacteria preparation freeze dried powder and aqueous pharmaceutical
Experiment purpose: compare Sargassum protein inhibiting-bacteria preparation freeze dried powder and the stability of solution under different temperatures.
Experimental technique: the extraction concentrate of getting an amount of Sargassum protein; Use contain sucrose and trehalose pH7.5 the 0.06mol/L phosphate buffer quantitatively dilution process Sargassum protein content be 500 μ g/ml solution (sucrose and trehalose be by weight the 1:1 proportioning, sucrose and trehalose account for the solution gross mass 5%).Get above-mentioned solution 1ml and be respectively charged in the cillin bottle, a part is used as the solution sample, and another part is processed freeze-dried powder through freeze drying, uses as the freeze dried powder sample.Respectively Sargassum protein solution sample and freeze dried powder sample are placed in 4 ℃ ,-20 ℃ environment, results of regular determination content, result such as below table 1:
Experimental result shows that the Sargassum protein solution was preserved 3 months in 4 ℃ of environment, in-20 ℃ of environment, preserve and can not survey content in 6 months; And freeze dried powder is preserved reduction by 30.4% in 24 months in 4 ℃ of environment, in-20 ℃ of environment, preserves 24 months stable contents.Can find out that Sargassum protein freeze dried powder of the present invention can preserve 2 years in-20 ℃ of environment, satisfy the storage of Sargassum protein inhibiting-bacteria preparation product and the requirement of application fully.
Experimental example 2The Sargassum protein inhibiting-bacteria preparation is the fungistatic effect experiment in the red bayberry preservation and freshness
Experimental technique: the Sargassum protein inhibiting-bacteria preparation 1g that takes the present invention's preparation is dissolved in the 1000ml pure water, and Sargassum protein concentration is about 0.02mg/ml, is sprayed on the new arbutus with the aerosol sprinkler; Other gets the commercially available mould azoles 1g of pressing down and is dissolved in the 1000ml pure water, and new arbutus was soaked wherein 2 minutes, pulls the back storage that drains away the water out, compares with the red bayberry of not placing any medicament.All experiments all place 4 ℃ of environment preservation and freshnesses, results of regular determination red bayberry SC sum content, and result such as table 2 are with shown in Figure 1.
Experimental result shows, can detect total plate count in control group storage under 4 ℃ of storage environments after 3 days, preserves to rot basically in 10 days and can't detect.Use Sargassum protein inhibiting-bacteria preparation and the red bayberry that presses down mould azoles can detect total plate count in storage after 10 and 8 days respectively, estimation is to press down to cause the red bayberry water content higher after mould azoles soaks, suitable bacterium colony development, some effects but the fungistatic effect of mould azoles.Can find out that Sargassum protein inhibiting-bacteria preparation of the present invention not only can reach the close effect of use chemical agent, and avoid chemical residual, make more consumers can eat red bayberry fresh fruit more relievedly.
Although the inventor has done comparatively detailed elaboration to technical scheme of the present invention and has enumerated; Be to be understood that; For the those skilled in the art in this area, be obvious to the replacement scheme that the foregoing description modifies and/or flexible perhaps employing is equal to, all can not break away from the essence of spirit of the present invention; The term that occurs among the present invention is used for can not being construed as limiting the invention the elaboration of technical scheme of the present invention and understanding.
Claims (2)
1. the application of Sargassum protein inhibiting-bacteria preparation in Waxberry preservation is characterized in that,
Described Sargassum protein inhibiting-bacteria preparation is for being mixed by following weight percentages after the freeze dried powder that obtains is handled in freeze drying:
Sargassum protein concentrate 1-50%,
Sucrose and trehalose 1-5%,
The phosphate buffer surplus,
Described Sargassum protein concentrate be fresh sheep dwell dish through the phosphate buffer lixiviate, filter, add solid (NH
4)
2SO
4Deposition, dialysis and PEG20000 concentrate and make, and obtain as follows:
(1) break process: get the fresh sheep dish of dwelling, the phosphate buffer that adds the 0.06mol/L of ice cube and pH 7.5 carries out fragmentation, and dwell dish and phosphate buffer of sheep is the mass ratio of 1:5～20,
(2) lixiviate, filtration, deposition, dialysis and concentration: the product after the break process is in 4 ℃ of following lixiviate 2h; With the coarse filtration of Sargassum protein leaching liquor, remove slag and get filtrating then, the protein extract after the coarse filtration is centrifugal in refrigerated centrifuge; Get supernatant, add solid (NH
4)
2SO
4Get 50% (NH
4)
2SO
4Deposition; The phosphate buffer dissolution precipitation of getting the 0.06mol/L of pH 7.5 is packed in the bag filter, dialyses in the phosphate buffer, and each dialysis time>=6h changes liquid number of times>=4 time; After dialysis is accomplished, concentrate with PEG20000 again, obtain the Sargassum protein concentrate;
The addition of described ice cube is as the criterion between 2 ~ 6 ℃ with the temperature in the control shattering process;
In the described freeze drying: the cold-trap precooling is to-35 ℃, and vacuum is 30～60Pa, and the time that freeze drying is handled is 8～9h;
Described Sargassum protein concentrate is preserved subsequent use down in-25 ℃～-10 ℃.
2. the application of Sargassum protein inhibiting-bacteria preparation according to claim 1 in Waxberry preservation is characterized in that described application is operated as follows:
Sargassum protein inhibiting-bacteria preparation 1g is dissolved in the 500-2500ml pure water, is sprayed on then on the new arbutus and gets final product.
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| Title |
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| 刘振宇等.海藻蛋白质提取物对香蕉炭疽菌的抑制作用.《福建农林大学学报(自然科学版)》.2006,第35卷(第1期),21-23. * |
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